Prism version 7.0a

Image analysis of cell migration counts and fungi growth was analyzed by hand using Image J software. Neutrophils in each chamber were counted every 15 min for the first 2 h of the experiment and then every hour for the remaining 16 h. Percentage of conidia to convert to hyphal growth was measured by counting conidia loaded per chamber before neutrophils or WBCs are loaded into the chamber and counting numbers of these conidia that grow hyphae by 18 h. Fungal growth velocity was calculated using Image J. For experiments with neutrophils from transplant patients, the 16 chambers in each device (n = 3) were analyzed for at least three different healthy donors. Data was analyzed for statistical significance using paired two-tailed t-tests. For zebrafish experiments, data was tested for normality using the D'Agostino & Pearson normality test. Normally distributed data was analyzed using two-tailed unpaired t-tests for pairwise comparisons, or ordinary one-way ANOVA with Tukey's multiple comparisons test. Non-normal data was compared using Kruskal-Wallis test for multiple comparisons. All statistical analysis was performed using Prism Version 7.0a (GraphPad).

Receptor stimulation protocols included ten sweeps of 25 s in duration, each of which consisted of two consecutive ACh pulses. A ‘desensitizing’ pulse of 3 s in duration was followed by a 50 ms ‘test’ pulse. The interval between pulses increased with each sweep from 300 ms to 7 s until full recovery from desensitization (Fig. 1A and B). Desensitization was analysed by fitting one or two exponential components to the current decay phase during the application of the desensitizing pulse. For better comparison of desensitization between groups, weighted current decay time constants were calculated according to: (τ₁ × A₁ + τ₂ × A₂)/(A₁ × A₂), where τ_x is the decay time constant for a particular component of the curve and A_x is the amplitude of the corresponding component. Recovery from desensitization was analysed as the ratio between peak current amplitudes of the test and desensitizing pulse in ten sweeps plotted against the interval between desensitizing and test pulse. Recovery time constants were determined by fitting one or two exponential components to the data points. Clampfit, version 10.5 (Molecular Devices) or Prism, version 7.0a (GraphPad Software Inc., San Diego, CA, USA) were used for exponential curve fitting and current decay or recovery time constant calculation, respectively.

Pairwise Student’s t-test was performed to check significance and denoted by one, two and three asterisks indicating p-value < 0.05, <0.01 and 0.001, respectively, for gene-expression related experiment. GraphPad PRISM (version 7.0a) https://www.graphpad.com/scientific-software/prism/ (accessed on 16 May 21) software was used to perform statistical analysis and for making the graphs. Adobe Illustrator CC version 2019, https://www.adobe.com/in/products/illustrator.html (accessed on 12 June 21) software was used to make final images.

Graphical and statistical analyses were performed using Prism version 7.0a (GraphPad Software). Kaplan-Meier curve analysis and log-rank tests were performed to reveal the impact of the different mutations, the treatment modalities, and several clinical variables on survival. p values between the different periods of treatment were determined using a non parametric unpaired t-test.